Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural AnalysesNitin Shirole 1, Sreeram Balasubramanian 1, Charles Yanofsky 2, Luis Cruz-Vera 1
1Department of Biological Sciences, University of Alabama Huntsville, 2Department of Biology, Stanford University
A major impediment to biochemical analyses of ribosomes containing nascent peptidyl-tRNAs has been the presence of other ribosomes in the same samples, ribosomes not involved in the translation of the specific mRNA sequence being analyzed. We developed a simple methodology to purify, exclusively, the ribosomes containing the nascent peptidyl-tRNA of interest.
